Method of preparing glucosylceramide synthase inhibitors

ABSTRACT

The invention relates to a method of preparing inhibitors of glucosylceramide synthase (GCS) useful for the treatment metabolic diseases, such as lysosomal storage diseases, either alone or in combination with enzyme replacement therapy, and for the treatment of cancer.

CROSS REFERENCE TO RELATED APPLICATION

This application is a continuation application of 371 National PhaseEntry application Ser. No. 14/776,443, filed Sep. 14, 2015 which claimsbenefit to International Application No. PCT/US2014/025384, filed Mar.13, 2014, which claims the benefit under 35 U.S.C. § 119 of U.S.Provisional Application 61/791,913, filed Mar. 15, 2013, all of whichare hereby incorporated by reference in their entirety.

BACKGROUND OF THE INVENTION

The invention relates to a method of preparing inhibitors ofglucosylceramide synthase (GCS) useful for the treatment metabolicdiseases, such as lysosomal storage diseases, either alone or incombination with enzyme replacement therapy, and for the treatment ofcancer.

Glucosylceramide synthase (GCS) is a pivotal enzyme which catalyzes theinitial glycosylation step in the biosynthesis of glucosylceramide-baseglycosphingolipids (GSLs) namely via the pivotal transfer of glucosefrom UDP-glucose (UDP-Glc) to ceramide to form glucosylceramide. GCS isa transmembrane, type III integral protein localized in the cis/medialGolgi. Glycosphingolipids (GSLs) are believed to be integral for thedynamics of many cell membrane events, including cellular interactions,signaling and trafficking. Synthesis of GSL structures has been shown(see, Yamashita et al., Proc. Natl. Acad. Sci. USA 1999, 96(16),9142-9147) to be essential for embryonic development and for thedifferentiation of some tissues. Ceramide plays a central role insphingolipid metabolism and downregulation of GCS activity has beenshown to have marked effects on the sphingolipid pattern with diminishedexpression of glycosphingolipids. Sphingolipids (SLs) have abiomodulatory role in physiological as well as pathologicalcardiovascular conditions. In particular, sphingolipids and theirregulating enzymes appear to play a role in adaptive responses tochronic hypoxia in the neonatal rat heart (see, El Alwanit et al.,Prostaglandins & Other Lipid Mediators 2005, 78(1-4), 249-263).

GCS inhibitors have been proposed for the treatment of a variety ofdiseases (see for example, WO2005068426). Such treatments includetreatment of glycolipid storage diseases (e.g., Tay Sachs, Sandhoffs,GM2 Activator deficiency, GM1 gangliosidosis and Fabry diseases),diseases associated with glycolipid accumulation (e.g., Gaucher disease;Miglustat (Zavesca), a GCS inhibitor, has been approved for therapy intype 1 Gaucher disease patients, see, Treiber et al., Xenobiotica 2007,37(3), 298-314), diseases that cause renal hypertrophy or hyperplasiasuch as diabetic nephropathy; diseases that cause hyperglycemia orhyperinsulemia; cancers in which glycolipid synthesis is abnormal,infectious diseases caused by organisms which use cell surfaceglycolipids as receptors, infectious diseases in which synthesis ofglucosylceramide is essential or important, diseases in which synthesisof glucosylceramide is essential or important, diseases in whichexcessive glycolipid synthesis occurs (e.g., atherosclerosis, polycystickidney disease, and renal hypertrophy), neuronal disorders, neuronalinjury, inflammatory diseases or disorders associated with macrophagerecruitment and activation (e.g., rheumatoid arthritis, Crohn's disease,asthma and sepsis) and diabetes mellitus and obesity (see, WO2006053043).

In particular, it has been shown that overexpression of GCS isimplicated in multi-drug resistance and disrupts ceramide-inducedapoptosis. For example, Turzanski et al., (Experimental Hematology 2005,33 (1), 62-72 have shown that ceramide induces apoptosis in acutemyeloid leukemia (AML) cells and that P-glycoprotein (p-gp) confersresistance to ceramide-induced apoptosis, with modulation of theceramide-glucosylceramide pathway making a marked contribution to thisresistance in TF-1 cells. Thus, GCS inhibitors can be useful fortreatment of proliferative disorders by inducing apoptosis in diseasedcells.

SUMMARY OF THE INVENTION

The present invention relates to a method of preparing a compound of theformula,

wherein:

n is 1, 2 or 3;

m is 1;

t is 0, 1 or 2;

y is 1 or 2;

z is 0, 1 or 2;

E is O;

X¹ is CR¹;

X² is O;

X³ is —NH;

X⁴ is CR⁴R⁵, CH₂CR⁴R⁵ or CH₂—(C₁-C₆) alkyl-CR⁴R⁵;

X⁵ is a direct bond, O, S, SO₂, CR⁴R⁵; (C₁-C₆)alkyl, (C₁-C₆)alkyloxy,(C₁-C₆)alkenyl, (C₁-C₆)alkenyloxy;

R is (C₆-C₁₂)aryl, (C₂-C₉)heteroaryl, (C₁-C₆)alkyl,(C₂-C₉)heteroaryl(C₁-C₆)alkyl;

R¹ is H, CN, (C₁-C₆)alkylcarbonyl, or (C₁-C₆)alkyl;

R² and R³ are each independently —H, (C₁-C₆)alkyl optionally substitutedby one or more substituents selected from the group consisting ofhalogen, (C₁-C₆)alkyl, (C₆-C₁₂)aryl, (C₂-C₉)heteroaryl,(C₁-C₆)alkyl(C₆-C₁₂)aryl, halo(C₆-C₁₂)aryl, and halo(C₂-C₉)heteroaryl,or optionally when X² is —NR² and X³ is —NR³, R² and R³ may be takentogether with the nitrogen atoms to which they are attached form anon-aromatic heterocyclic ring optionally substituted by with one ormore substituents selected from halogen, (C₁-C₆)alkyl, (C₆-C₁₂)aryl,(C₂-C₉)heteroaryl, (C₁-C₆)alkyl(C₆-C₁₂)aryl, halo(C₆-C₁₂)aryl, andhalo(C₂-C₉)heteroaryl;

R⁴ and R⁵ are independently selected from H, (C₁-C₆)alkyl, or takentogether with the carbon to which they are attached to form a spiro(C₃-C₁₀)cycloalkyl ring or spiro (C₃-C₁₀)cycloalkoxy ring;

R⁶ is —H, halogen, —CN, (C₆-C₁₂)aryl, (C₆-C₁₂)aryloxy, (C₁-C₆)alkyloxy;(C₁-C₆)alkyl optionally substituted by one to four halo or (C₁-C₆)alkyl;

A¹ is (C₂-C₆)alkynyl; (C₃-C₁₀)cycloalkyl, (C₆-C₁₂)aryl,(C₂-C₉)heteroaryl, (C₂-C₉)heterocycloalkyl orbenzo(C₂-C₉)heterocycloalkyl optionally substituted with one or moresubstituents selected from the group consisting of halo, (C₁-C₆)alkyloptionally substituted by one to three halo; (C₁-C₆)alkenyl, amino,(C₁-C₆)alkylamino, (C_(r) C₆)dialkylamino, (C₁-C₆)alkoxy, nitro, CN,—OH, (C₁-C₆)alkyloxy optionally substituted by one to three halo;(C₁-C₆)alkoxycarbonyl, and (C₁-C₆) alkylcarbonyl;

A² is H, (C₃-C₁₀)cycloalkyl, (C₆-C₁₂)aryl, (C₂-C₉)heteroaryl,(C₂-C₉)heterocycloalkyl or benzo(C₂-C₉)heterocycloalkyl optionallysubstituted with one or more substituents selected from the groupconsisting of halo, (C₁-C₆)alkyl optionally substituted by one to threehalo; (C₁-C₆)alkylenyl, amino, (C₁-C₆) alkylamino, (C₁-C₆)dialkylamino,(C₁-C₆)alkoxy, O(C3-C6 cycloalkyl), (C₃-C₆) cycloalkoxy, nitro, CN, OH,(C₁-C₆)alkyloxy optionally substituted by one to three halo; (C₃-C₆)cycloalkyl, (C₁-C₆) alkoxycarbonyl, (C₁-C₆) alkylcarbonyl, (C₁-C₆)haloalkyl;

with the proviso that the sum of n+t+y+z is not greater than 6;comprising reacting the compound of Formula II

with the compound of Formula III

wherein n, t, y, z, X⁴, A¹, X⁵ and A² are as defined above.

The present invention relates to a method of preparing a compound of theformula,

wherein:

n is 1, 2 or 3;

m is 1;

t is 0, 1 or 2;

y is 1 or 2;

z is 0, 1 or 2;

E is O;

X¹ is CR¹;

X² is O;

X³ is —NH;

X⁴ is CR⁴R⁵, CH₂CR⁴R⁵ or CH₂—(C₁-C₆) alkyl-CR⁴R⁵;

X⁵ is a direct bond, O, S, SO₂, CR⁴R⁵; (C₁-C₆)alkyl, (C₁-C₆)alkyloxy,(C₁-C₆)alkenyl, (C₁-C₆)alkenyloxy;

R is (C₆-C₁₂)aryl, (C₂-C₉)heteroaryl, (C₁-C₆)alkyl,(C₂-C₉)heteroaryl(C₁-C₆)alkyl;

R¹ is H, CN, (C₁-C₆)alkylcarbonyl, or (C₁-C₆)alkyl;

R² and R³ are each independently —H, (C₁-C₆)alkyl optionally substitutedby one or more substituents selected from the group consisting ofhalogen, (C₁-C₆)alkyl, (C₆-C₁₂)aryl, (C₂-C₉)heteroaryl,(C₁-C₆)alkyl(C₆-C₁₂)aryl, halo(C₆-C₁₂)aryl, and halo(C₂-C₉)heteroaryl,or optionally when X² is —NR² and X³ is —NR³, R² and R³ may be takentogether with the nitrogen atoms to which they are attached form anon-aromatic heterocyclic ring optionally substituted by with one ormore substituents selected from halogen, (C₁-C₆)alkyl, (C₆-C₁₂)aryl,C₁₂)aryl, (C₂-C₉)heteroaryl, (C₁-C₆)alkyl(C₆-C₁₂)aryl, halo(C₆-C₁₂)aryl,and halo(C₂-C₉)heteroaryl;

R⁴ and R⁵ are independently selected from H, (C₁-C₆)alkyl, or takentogether with the carbon to which they are attached to form a spiro(C₃-C₁₀)cycloalkyl ring or spiro (C₃-C₁₀)cycloalkoxy ring;

R⁶ is —H, halogen, —CN, (C₆-C₁₂)aryl, (C₆-C₁₂)aryloxy, (C₁-C₆)alkyloxy;(C₁-C₆)alkyl optionally substituted by one to four halo or (C₁-C₆)alkyl;

A¹ is (C₂-C₆)alkynyl; (C₃-C₁₀)cycloalkyl, (C₆-C₁₂)aryl,(C₂-C₉)heteroaryl, (C₂-C₉)heterocycloalkyl orbenzo(C₂-C₉)heterocycloalkyl optionally substituted with one or moresubstituents selected from the group consisting of halo, (C₁-C₆)alkyloptionally substituted by one to three halo; (C₁-C₆)alkenyl, amino,(C₁-C₆)alkylamino, (C_(r) C₆)dialkylamino, (C₁-C₆)alkoxy, nitro, CN,—OH, (C₁-C₆)alkyloxy optionally substituted by one to three halo;(C₁-C₆)alkoxycarbonyl, and (C₁-C₆) alkylcarbonyl;

A² is H, (C₃-C₁₀)cycloalkyl, (C₆-C₁₂)aryl, (C₂-C₉)heteroaryl,(C₂-C₉)heterocycloalkyl or benzo(C₂-C₉)heterocycloalkyl optionallysubstituted with one or more substituents selected from the groupconsisting of halo, (C₁-C₆)alkyl optionally substituted by one to threehalo; (C₁-C₆)alkylenyl, amino, (C₁-C₆) alkylamino, (C₁-C₆)dialkylamino,(C₁-C₆)alkoxy, O(C3-C6 cycloalkyl), (C₃-C₆) cycloalkoxy, nitro, CN, OH,(C₁-C₆)alkyloxy optionally substituted by one to three halo; (C₃-C₆)cycloalkyl, (C₁-C₆) alkoxycarbonyl, (C₁-C₆) alkylcarbonyl, (C₁-C₆)haloalkyl;

with the proviso that the sum of n+t+y+z is not greater than 6;comprising reacting the compound of Formula IV

with a compound of Formula III

wherein n, t, y, z, X⁴, A¹, X⁵ and A² are as defined above.The present invention relates to a method of preparing a compound of theformula,

wherein:

n is 1, 2 or 3;

m is 1;

t is 0, 1 or 2;

y is 1 or 2;

z is 0, 1 or 2;

E is O;

X¹ is CR¹;

X² is O;

X³ is —NH;

X⁴ is CR⁴R⁵, CH₂CR⁴R⁵ or CH₂—(C₁-C₆) alkyl-CR⁴R⁵;

X⁵ is a direct bond, O, S, SO₂, CR⁴R⁵; (C₁-C₆)alkyl, (C₁-C₆)alkyloxy,(C₁-C₆)alkenyl, (C₁-C₆)alkenyloxy;

R is (C₆-C₁₂)aryl, (C₂-C₉)heteroaryl, (C₁-C₆)alkyl,(C₂-C₉)heteroaryl(C₁-C₆)alkyl;

R¹ is H, CN, (C₁-C₆)alkylcarbonyl, or (C₁-C₆)alkyl;

R² and R³ are each independently —H, (C₁-C₆)alkyl optionally substitutedby one or more substituents selected from the group consisting ofhalogen, (C₁-C₆)alkyl, (C₆-C₁₂)aryl, (C₂-C₉)heteroaryl,(C₁-C₆)alkyl(C₆-C₁₂)aryl, halo(C₆-C₁₂)aryl, and halo(C₂-C₉)heteroaryl,or optionally when X² is —NR² and X³ is —NR³, R² and R³ may be takentogether with the nitrogen atoms to which they are attached form anon-aromatic heterocyclic ring optionally substituted by with one ormore substituents selected from halogen, (C₁-C₆)alkyl, (C₆-C₁₂)aryl,(C₂-C₉)heteroaryl, (C₁-C₆)alkyl(C₆-C₁₂)aryl, halo(C₆-C₁₂)aryl, andhalo(C₂-C₉)heteroaryl;

R⁴ and R⁵ are independently selected from H, (C₁-C₆)alkyl, or takentogether with the carbon to which they are attached to form a spiro(C₃-C₁₀)cycloalkyl ring or spiro (C₃-C₁₀)cycloalkoxy ring;

R⁶ is —H, halogen, —CN, (C₆-C₁₂)aryl, (C₆-C₁₂)aryloxy, (C₁-C₆)alkyloxy;(C₁-C₆)alkyl optionally substituted by one to four halo or (C₁-C₆)alkyl;

A¹ is (C₂-C₆)alkynyl; (C₃-C₁₀)cycloalkyl, (C₆-C₁₂)aryl,(C₂-C₉)heteroaryl, (C₂-C₉)heterocycloalkyl orbenzo(C₂-C₉)heterocycloalkyl optionally substituted with one or moresubstituents selected from the group consisting of halo, (C₁-C₆)alkyloptionally substituted by one to three halo; (C₁-C₆)alkenyl, amino,(C₁-C₆)alkylamino, (C₁-C₆)dialkylamino, (C₁-C₆)alkoxy, nitro, CN, —OH,(C₁-C₆)alkyloxy optionally substituted by one to three halo;(C₁-C₆)alkoxycarbonyl, and (C₁-C₆) alkylcarbonyl;

A² is H, (C₃-C₁₀)cycloalkyl, (C₆-C₁₂)aryl, (C₂-C₉)heteroaryl,(C₂-C₉)heterocycloalkyl or benzo(C₂-C₉)heterocycloalkyl optionallysubstituted with one or more substituents selected from the groupconsisting of halo, (C₁-C₆)alkyl optionally substituted by one to threehalo; (C₁-C₆)alkylenyl, amino, (C₁-C₆) alkylamino, (C₁-C₆)dialkylamino,(C₁-C₆)alkoxy, O(C3-C6 cycloalkyl), (C₃-C₆) cycloalkoxy, nitro, CN, OH,(C₁-C₆)alkyloxy optionally substituted by one to three halo; (C₃-C₆)cycloalkyl, (C₁-C₆) alkoxycarbonyl, (C₁-C₆) alkylcarbonyl, (C₁-C₆)haloalkyl;

with the proviso that the sum of n+t+y+z is not greater than 6;comprising reacting the compounds of Formula II and Formula IV

with a compound of Formula III

wherein n, t, y, z, X⁴, A¹, X⁵ and A² are as defined above.

The present invention further relates to a method wherein n is 1; t is0; y is 1 and z is 1.

The present invention further relates to a method wherein X⁴ is CR⁴R⁵.

The present invention further relates to a method wherein R⁴ and R⁵ areeach methyl.

The present invention further relates to a method wherein A¹ is(C₂-C₉)heteroaryl.

The present invention further relates to a method wherein A¹ isthiophene, thiazole, isothiazole, furane, oxazole, isoxazole, pyrrole,imidazole, pyrazole, triazole, pyridine, pymiridine, pyridazine, indole,benzotiazole, benzoisoxazole, benzopyrazole, benzoimidazole, benzofuran,benzooxazole or benzoisoxazole.

The present invention further relates to a method wherein A¹ isthiazole.

The present invention further relates to a method wherein R⁶ is H.

The present invention further relates to a method wherein X⁵ is a directbond.

The present invention further relates to a method wherein A² is(C₆-C₁₂)aryl.

The present invention further relates to a method wherein A² is phenyl.

The present invention further relates to a method wherein the phenylgroup is substituted by halo.

The present invention further relates to a method wherein the halo groupis fluoro.

The present invention further relates to a method wherein R¹ ishydrogen.

The present invention further relates to a method including reacting thecompound of Formula V

with imidazole to form the compound of Formula II

wherein X⁴, A¹, X⁵ and A² are as defined above.

The present invention further relates to a method including heating toreflux the compound of Formula V

to form the compound of Formula IV

wherein X⁴, A¹, X⁵ and A² are as defined above.

The present invention further relates to a method including reacting,while heating to reflux, the compound of Formula V

with imidazole to form the compounds of Formula II and Formula IV

wherein X⁴, A¹, X⁵ and A² are as defined above.

The present invention further relates to a method including reacting thecompound of Formula VI

with N, N′-carbonyldiimidazole to form the compound of Formula V

wherein X⁴, A¹, X⁵ and A² are as defined above.

The present invention further relates to a method including reacting thecompound of Formula VII

with N, N′-carbonyldiimidazole and hydroxylamine to form the compound ofFormula VI

wherein X⁴, A¹, X⁵ and A² are as defined above.

The present invention relates to a method of preparing the compound ofFormula VIII

comprising reacting a compound of Formula IX

with quinuclidinol.

The present invention relates to a method of preparing the compound ofFormula VIII

comprising reacting a compound of Formula X

with quinuclidinol.

The present invention relates to a method of preparing the compound ofFormula VIII

comprising reacting a compounds of Formula IX and Formula X

with quinuclidinol.

The present invention further relates to a method including reacting thecompound of Formula XI

with imidazole to form the compound of Formula IX

The present invention further relates to a method including heating toreflux the compound of Formula XI

to form the compound of Formula X

The present invention further relates to a method including reacting,while heating to reflux, the compound of Formula XI

with imidazole to form the compounds of Formula IX and Formula X

The present invention further relates to a method including reacting thecompound of Formula XII

with N, N′-carbonyldiimidazole to form the compound of Formula XI

The present invention further relates to a method including reacting thecompound of Formula XIII

with N, N′-carbonyldiimidazole and hydroxylamine to form the compound ofFormula XII

The present invention further relates to a method including reacting thecompound of Formula XIV

with potassium tert-butoxide and methyl iodine followed by reacting theethyl ester so formed with lithium hydroxide to form the compound ofFormula XIII

The present invention relates to a compound of Formula XII

The present invention relates to a compound of Formula XI

The present invention relates to a compound of Formula IX

DETAILED DESCRIPTION OF THE INVENTION

In reaction 1 of Scheme 1, the carboxylic acid compound of Formula VIIis converted to the corresponding hydroxamic acid compound of Formula VIby reacting VII with N, N′-carbonyldiimidazole (i.e. CDI) in a polaraprotic solvent, such as tetrahydrofuran (THF). The solution is stirredat a temperature between about −5° C. to about 25° C., preferable about20° C., for a time period between about 5 minutes to about 30 minutes,preferably about 10 to 15 minutes. The resulting solution mixture isallowed to warm to room temperature and stirred for an additional timeperiod between about 30 minutes to about 2 hours, preferably about 1hour. Hydroxylamine is then added to the solution mixture at atemperature between about −5° C. to about 10° C., preferable about 3° C.The resulting reaction mixture is stirred under inert atmosphere (i.e.,nitrogen) for a time period between about 5 min to about 8 hours,preferably about 10 min.

In reaction 2 of Scheme 1, the hydroxamic acid compound of Formula VI isconverted to the corresponding compound of Formula V by the addition ofN, N′-carbonyldiimidazole to a solution of VI in toluene under inertatmosphere (i.e., nitrogen) and stirred for a time period between about30 minutes to about 4 hours, preferably about 2.5 hours.

In reaction 3 of Scheme 1, the compound of Formula V is converted to thecorresponding compounds of Formula II and Formula IV by reacting V withimidazole in the presence of a aprotic solvent, such as toluene. Thereaction mixture is heated to reflux for a time period between about 4hours to about 28 hours, preferable about 6 hours.

In reaction 4 of Scheme 1, a mixture of the compounds of Formula II andFormula IV (or each intermediate separately) is converted to thecorresponding compound of Formula I by reacting II and IV with(S)-(+)-quinuclidinol in the presence of a aprotic solvent, such astoluene. The reaction mixture is heated to reflux for a time periodbetween about 12 hours to about 24 hours, preferable about 18 hours.

Preparation A

To 4-Fluorophenylthioamide (50.35 g, 1 eq.) was added 8.6 weight volumesof 200 proof ethanol (based on thioamide) (430 mL) and ethyl4-chloroacetoacetate (68.2 g, 1.1 eq.). The mixture was place under anitrogen atmosphere. It was heated under reflux for 5 h and allowed tocool to room temperature. The solution was concentrated to an oil andTBME (10 volumes, 500 mL) and 6 volumes of saturated NaHCO3 (300 mL)added. The aqueous layer was back extracted with 5 volumes (250 mL) ofTBME. The combined organic layer was washed with water and thenconcentrated to an oil and then dried to a solid. The product wascrystallized from 3 weight volumes of hot hexanes. Yield 89% Product98.7% pure by HPLC (area %).

Example 1 (S)-Quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate

Step 1: Dimethylation with Methyl Iodide

Procedure:

In a 100 L reactor was added tetrahydrofuran (THF, 28.4 Kg) andpotassium tert-butoxide (MW 112.21, 2.28 Kg g, 4.0 equiv.,). Thismixture was cooled to 0-2° C. (internal temperature). The starting ester(MW 265.3, 2.0 Kg, 1.0 equiv.) was dissolved in THF (4 L) andtransferred to the reactor over a period of 10-60 min, keeping theinternal temperature below 10° C. during the addition. The reactionmixture was stirred at 3-9° C. for 15-60 min. A solution of methyliodide (MW 141.94, 1.88 L, 4.0 equiv.) in THF (4.8 L) was added to thereactor over 30-120 min keeping the internal temperature below 10° C. Asolution of NaCl (2.0 Kg) in water (14 L) was added over 10 min and themixture was stirred for at least 10 min more. The reaction was madeacidic by the addition of 1 M HCl (˜1.44 L). The layers were separatedand the aqueous layer was back extracted with THF (6.2 kg). The combinedorganic layers were vacuum distilled to ˜16 l. This THF solution of theStep 1 product was used in the next reaction.

Step 2: Hydrolysis of the Ethyl Ester with LiOH Monohydrate

Procedure:

To the ester in THF was added a solution of LiOH.H₂O (MW 41.96, 0.695Kg, 2.2 equiv.) in water (9.3 L) was added. The mixture was heated atreflux for 8-16 hours. After the reactions was judge complete by HPLC,water (12 L) was added and the mixture was vacuum distilled to ˜16 L.TBME (5.9 kg) was added and after stirring the layers were separated.The aqueous layer containing the product was washed a second time withTBME (5.9 Kg). TBME was added to the aqueous layer and the mixture wasmade acidic (pH≤3) by the addition of 5 M HCl (˜3.67 Kg). The layerswere separated and the aqueous layer was extracted a second time withTBME (4.5 Kg). Heptane (15 Kg) was added to the combined organic layersand the mixture was vacuum distilled to 16 L. After heating and coolingto 5-25° C. and stirring for at least 3 h, the product was filtered,washed with heptane, and vacuum dried. Yield 85.8% (2.15 Kg) HPLC purity(area %) 99.72%

Reaction 1: Formation of Hydroxamic Acid with NH₂OH

Procedure:

To a 100 L reactor was added THF (14.2 Kg) and N, N′-carbonyldiimidazole(CDI; MW 162.15, 1.34 Kg, 1.1 equiv.). The acid from reaction 2 (2.0 Kg,1.0 equiv) dissolved in THF (4 L) was added over 15-20 min. The mixturewas stirred at room temperature for 2.5-3 h. The reaction was cooled to0-3° C. Aqueous hydroxylamine (50% aqueous; 1.7 L, 4.0 equiv.) was addedover 5-15 min keeping the internal temperature less than 18° C. Afterthe addition was complete, the layers were separated and the organiclayer was washed with water (12 Kg) and a solution of sodium chloride(2.0 Kg) in water (12 L). The separated organic layer was vacuum distillto ˜16 l. Toluene (13.8 Kg) was added and the mixture was again vacuumdistilled to ˜16 L. Heptane (11 kg) was added and the mixture wasstirred at room temperature for at least 16 h. The resulting solid wasfiltered, washed with heptane (11 Kg) and vacuum dried at roomtemperature. The yield was 1.58 Kg (74.8%).

Reaction 2: Conversion of Hydroxamic Acid to a Dioxazolone

Procedure:

Toluene (17.3 Kg) and the hydroxamic acid from, reaction 1 (MW 280.32,2.0 Kg) was transferred to a 100 L reactor. After stirring at roomtemperature for at least 15 min carbonyl diimidazole CDI (MW 162.15,1.27 Kg, 1.1 equiv.) was added. The mixture was stirred at roomtemperature for 1-4 h until the reaction was judge complete by HPLC.

Reaction 3 Conversion of the Dioxazolone to a Mixture of the ImidazoleUrea and Isocyanate

Procedure:

The solution of the dioxazolone (reaction 2) was heated at 60° C. for6-16 hours to complete the conversion to a mixture of the isocyanate andimidazole urea as judge by HPLC analysis.

Reaction 4: Final Conversion to the Carbamate

Procedure:

(S)-(+)-3-quinuclidinol (1.14 Kg, 1.18 equiv.) was added to the mixtureof the isocyanate and imidazole urea toluene solution (reaction 3) andthe solution was heated at 100-110° C. for 18-28 h. Toluene (8.6 Kg) wasadded to the reaction and the mixture was washed twice with water (20Kg). The product was removed from the organic layer with two extractionsof aqueous 1M HCl (19.7 Kg). Isopropyl acetate (34.8 Kg) was added tothe combined acidic aqueous layers. The mixture was cooled to 5-10° C.and 10M aqueous NaOH (5.3 Kg) was added. The layers were separated andthe organic layer was vacuum distilled to ˜16 L. Heptane (21.4 Kg) wasadded to the remaining isopropyl acetate solution and again the solutionwas distilled to 16 L. the resulting suspension was stirred for at least4 h. The product was filtered, washed with heptane (13.7 Kg) and vacuumdried at room temperature. The yield was 2.3 Kg (82.8% yield). HPLCpurity (Area %) 99.7%.

¹H NMR (400 MHz, CDCl₃) δ8.04-7.83 (m, 2H), 7.20-6.99 (m, 3H), 5.53 (s,1H), 4.73-4.55 (m, 1H), 3.18 (dd, J=14.5, 8.4 Hz, 1H), 3.05-2.19 (m,5H), 2.0-1.76 (m, 11H). ¹³C NMR (100 MHz, CDCl₃) δ166.38, 165.02,162.54, 162.8-155.0 (d, C—F), 130.06, 128.43, 128.34, 116.01, 115.79,112.46, 71.18, 55.70, 54.13, 47.42, 46.52, 27.94, 25.41, 24.67, 19.58.

Example 2 (S)-Quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate

2-(2-(4-fluorophenyl)thiazol-4-yl)-2-methylpropanoic acid (1 g) anddiisopropylethyl amine (0.57 ml) were dissolved in toluene and stirredat 110° C. under N2. DPPA (0.9 ml) was added dropwise. The mixture wasstirred for 3 hours at 110° C. to complete the conversion of the acetylazide and isocyanate. Quinuclidin-3-ol (0.72 g) was added and stirredfor 18 hours. The result mixture was diluted with toluene (50 ml) andwashed with saturated sodium bicarbonate solution. The organic layer wasconcentrated to oil. Product of quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate was purified bycrystallization from EtOAc (0.6 g).

The invention claimed is:
 1. A method of preparing the compound of Formula VIII

comprising reacting a compound of Formula IX

with quinuclidinol.
 2. A method according to claim 1, further including reacting the compound of Formula XI

with imidazole to form the compound of Formula IX


3. A method according to claim 1, further including heating to reflux the compound of Formula XI

to form the compound of Formula X


4. A method according to claim 1, further including reacting, while heating to reflux, the compound of Formula XI

with imidazole to form the compounds of Formula IX and Formula X


5. A method according to claim 2, further including reacting the compound of Formula XII

with N, N′-carbonyldiimidazole to form the compound of Formula XI


6. A method according to claim 5, further including reacting the compound of Formula XIII

with N, N′-carbonyldiimidazole and hydroxylamine to form the compound of Formula XII


7. A method according to claim 6, further including reacting the compound of Formula XIV

with potassium tert-butoxide and methyl iodine followed by reacting the ethyl ester so formed with lithium hydroxide to form the compound of Formula XIII. 